RESUMO
A peptide self-assembled monolayer (SAM) was designed to bind His-tagged biomolecules for surface plasmon resonance (SPR) bioanalysis, which was applied for the determination of K(d) for small ligand screening against CD36. Nonspecific adsorption could be minimized using penta- and hexa-peptide monolayers. In particular, monolayers consisting of 3-mercaptopropionyl-leucinyl-histidinyl-aspartyl-leucinyl-histidinyl-aspartic acid (3-Mpa-LHDLHD) exhibited little (12 ng cm(-2)) nonspecific adsorption in crude serum. Modification of this peptide monolayer with Nα,Nα-bis(carboxymethyl)-L-lysine gave a surface competent for binding His-tagged proteins, as demonstrated using enzyme (human dihydrofolate reductase), protein/antibody and receptor (CD36) examples. Immobilization featured chelation of copper and the His-tagged protein by the peptide monolayer, which could be recycled by removing the copper using imidazole washes prior to reuse.
Assuntos
Antígenos CD36/metabolismo , Histidina/química , Peptídeos/química , Bibliotecas de Moléculas Pequenas/farmacologia , Ressonância de Plasmônio de Superfície/métodos , Adsorção , Sequência de Aminoácidos , Animais , Bovinos , Avaliação Pré-Clínica de Medicamentos/métodos , Histidina/metabolismo , Humanos , Proteínas Imobilizadas/química , Proteínas Imobilizadas/metabolismo , Ligantes , Peptídeos/metabolismo , Ligação Proteica , Tetra-Hidrofolato Desidrogenase/química , Tetra-Hidrofolato Desidrogenase/metabolismoRESUMO
[reaction: see text] Macrocyclic and fused bicyclic dipeptides are complementary motifs for mimicry of different types of beta-turn geometry. Macrocyclic dipeptide mimics have served as precursors for the synthesis of their bicyclic counterparts using electrophilic transannular cyclizations of 9- and 10-membered ring lactams 9-12 to form azabicyclo[4.3.0]- and -[5.3.0]alkanone amino esters 13-16.
Assuntos
Aminoácidos/síntese química , Compostos Bicíclicos Heterocíclicos com Pontes/síntese química , Dipeptídeos/química , Lactamas/química , Aminoácidos/química , Compostos Bicíclicos Heterocíclicos com Pontes/química , Catálise , Ciclização , Mimetismo Molecular , Estrutura MolecularRESUMO
The Rho signaling pathway regulates the cytoskeleton and motility and plays an important role in neuronal growth inhibition. Here we demonstrate that inactivation of Rho or its downstream target Rho-associated kinase (ROK) stimulated neurite growth in primary cells of cortical neurons plated on myelin or chondroitin sulfate proteoglycan substrates. Furthermore, treatment either with C3 transferase (C3) to inactivate Rho or with Y27632 to inhibit ROK was sufficient to stimulate axon regeneration and recovery of hindlimb function after spinal cord injury (SCI) in adult mice. Injured mice were treated with a single injection of Rho or Rho-associated kinase inhibitors delivered in a protein adhesive at the lesion site. Treated animals showed long-distance regeneration of anterogradely labeled corticospinal axons and increased levels of GAP-43 mRNA in the motor cortex. Behaviorally, inactivation of Rho pathway induced rapid recovery of locomotion and progressive recuperation of forelimb-hindlimb coordination. These findings provide evidence that the Rho signaling pathway is a potential target for therapeutic interventions after spinal cord injury.